title: Development of highly polymorphic EST-SSR markers and segregation in F₁ hybrid population of Vitis vinifera L.
abstract: The objectives of this investigation were to develop and validate the expressed sequence tag (EST)-simple sequence repeat (SSR) markers from large EST sequences, and to study the segregation and distribution of SSRs within two grapevine parental lines. In total, 94 F₁ lines crossed between "Early Rose" and "Red Globe" were studied. Approximately 2100 EST-SSR sequences of Vitis vinifera L. were searched for SSRs and analyzed for the design of polymerase chain reaction (PCR) primers amplifying the SSR-rich regions. Trinucleotide repeats were found to be the most abundant, followed by other nucleotide repeats. A total of 182 SSR primer pairs were first developed for the study on the parental polymorphism. Among the 182 SSR primers, 142 primer pairs (78%) could amplify the anticipated PCR products, among which only 52 primer pairs (36.62%) showed polymorphism between the two parents. These polymorphic bands were further surveyed among the 94 F₁ lines, and the results showed that a total of 162 bands were amplified, and 98 of them were polymorphic in both parents (60.86% polymorphism), with an average of 1.88 polymorphic DNA bands for each primer pair. After testing with the chi-square test, 33 of the clearly amplified polymorphic bands followed a 3:1 ratio, and 37 followed a 1:1 ratio. The rest showed distorted segregation ratios.
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing, China.
authors.forename: E
authors.initials: E
authors.lastname: Kayesh
authors.forename: Y Y
authors.initials: YY
authors.lastname: Zhang
authors.forename: G S
authors.initials: GS
authors.lastname: Liu
authors.forename: N
authors.initials: N
authors.lastname: Bilkish
authors.forename: X
authors.initials: X
authors.lastname: Sun
authors.forename: X P
authors.initials: XP
authors.lastname: Leng
authors.forename: J G
authors.initials: JG
authors.lastname: Fang
mesh_terms: D002874:Chromosome Mapping; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D020224:Expressed Sequence Tags; D005819:Genetic Markers; D018745:Genome, Plant; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D027843:Vitis / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018744:DNA, Plant; D005819:Genetic Markers